培养基优化以促进超高浓度发酵生产乙醇(英文)

An optimal medium (300 g·L-1 initial glucose) comprising 6.3 mmol·L-1 Mg2+, 5.0 mmol·L-1 Ca2+, 15.0 g·L-1 peptone and 21.5 g·L-1 yeast extract was determined by uniform design to improve very high gravity (VHG) ethanol fermentation, showing over 30% increase in final ethanol (from 13.1% to 17.1%, by volume), 29% decrease in fermentation time (from 84 to 60 h), 80% increase in biomass formation and 26% increase in glucose utilization. Experiments also revealed physiological aspects linked to the fermentation enhancements. Compared to the control, trehalose in the cells grown in optimal fermentation medium increased 17.9-, 2.8-, 1.9-, 1.8- and 1.9-fold at the fermentation time of 12, 24, 36, 48 and 60 h, respectively. Its sharp rise at the early stage of fermentation when there was a considerable osmotic stress suggested that trehalose played an important role in promoting fermentation. Meanwhile, at the identical five fermentation time, the plasma membrane ATPase activity of the cells grown in optimal medium was 2.3, 1.8, 1.6, 1.5 and 1.3 times that of the control, respectively. Their disparities in enzymatic activity became wider when the glucose levels were dramatically changed for ethanol production, suggesting this enzyme also contributed to the fermentation improvements. Thus, medium optimization for VHG ethanol fermentation was found to trigger the increased yeast trehalose accumulation and plasma membrane ATPase activity.     

酒精-沼气双发酵耦联工艺中丙酸对酿酒酵母酒精发酵的影响(英文)

Propanoic acid accumulated in an ethanol-methane coupled fermentation process affects the ethanol fermentation by Saccharomyces cerevisiae. The effects of propanoic acid on ethanol production were examined in cassava mash under different pH conditions. Final ethanol concentrations increased when undissociated propanoic acid was <30.0 mmol·L-1 . Propanoic acid, however, stimulated ethanol production, as much as 7.6% under proper conditions, but ethanol fermentation was completely inhibited when undissociated acid was >53.2 mmol·L-1 . Therefore, the potential inhibitory effect of propanoic acid on ethanol fermentation may be avoided by controlling the undissociated acid concentrations through elevated medium pH. Biomass and glycerol production decreased with propanoic acid in the medium, partly contributing to increased ethanol concentration.     

L-半胱氨酸浓度对氧化还原电位和生物产氢过程的影响(英文)

The effects of L-cysteine concentration on biohydrogen production by Enterobacterium Bacterium M580 were investigated in batch cultivation.The experimental results showed that L-cysteine could enhance the cell growth,hydrogen production rate and hydrogen yield when its concentration was less than 500 mg·L-1,while it had negative effects when its concentration was higher than 500 mg·L-1.The hydrogen production was the highest 1.29 mol·mol-1(H2/glucose) when 300 mg·L-1L-cysteine was added into the culture,and the yield was 9.4% higher than that in the control.The oxidation-reduction potential(ORP) ,which was influenced by L-cysteine,also affected hydrogen production.The ORP values were in the range-300 mV to-150 mV when the L-cysteine concentration was higher than 500 mg·L-1.Although the ORP in this range was favorable for hydrogen production,it was not suitable for the biomass growth.Hence,less hydrogen was produced.When the L-cysteine concentration was lower than 500 mg·L-1,the ORP was more suitable for both biomass growth and hydrogen production.In addition,at least 91%glucose was consumed when L-cysteine was added to the culture media,compared to the 97.37% consumption without L-cysteine added.     

短乳杆菌CGMCC1306pH和温度两阶段控制结合底物补加方式生产γ-氨基丁酸(英文)

Methods to optimize the production of gamma-aminobutyric acid （GABA） by Lactobacillus brevis CGMCC 1306 were investigated. Results indicated that cell growth was maximal at pH 5.0, while pH 4.5 was pref-erable to GABA formation. The optimal temperature for cell growth （35 °C） was lower than that for GABA forma-tion （40 °C）. In a two-stage pH and temperature control fermentation, cultures were maintained at pH 5.0 and 35 °C for 32 h, then adjusted to pH 4.5 and 40 °C, GABA production increased remarkably and reached 474.79 mmol·L-1 at 72 h, while it was 398.63 mmol·L-1 with one stage pH and temperature control process, in which cultivation con-ditions were constantly controlled at pH 5.0 and 35 °C. In order to avoid the inhibition of cell growth at higher L-monosodium glutamate （L-MSG） concentrations, the two-stage control fermentation with substrate feeding strat-egy was applied to GABA production, with 106.87 mmol （20 g） L-MSG supplemented into the shaking-flask at 32 h and 56 h post-inoculation separately. The GABA concentration reached 526.33 mmol·L-1 at 72 h with the fer-mentation volume increased by 38%. These results will provide primary data to realize large-scale production of GABA by L. brevis CGMCC 1306.     

Kinetics of Asymmetric Reduction of Phenylglyoxylic Acid to R-(－)-Mandelic Acid by Saccharomyces Cerevisiae FD11b

The kinetics of asymmetric production of R-(-)-mandelic acid (R-MA) from phenylglyoxylic acid (PGA)catalyzed by Saccharomyces cerevisiae sp. strain FD11b was studied by fed-batch cultures. The concentrations of glucose and PGA were controlled respectively with a dual feeding system. When the electron donor glucose was supplied at the rate of 0.0833mmol·gdw-1·h-1, the specific production rate (qp) and the enantiomeric excess of R-MAreached the maximum 0.353mmol·gdw-1·h-1 and 97.1％, respectively. The apparent reduction activity of yeast FD11b was obviously affected by both substrate PGA and product MA. The qp value reached the maximum 0.36-0.38mmol·gdw-1·h-1 when the PGA concentration was controlled between 25 and 35mmol·L-1. The obvious substrate inhibition of bioconversion was observed at the PGA concentrations higher than 40mmol· L-1. The accumulation of product MA also caused a severe feed-back inhibition for its production when the product concentration was above 60mmol· L-1. The kinetic model with the inhibition effect of both substrate and product was simulated by a computer-based least-square arithmatic. The established kinetic model was in good agreement with the experimental data.     

钠盐浓度对厌氧产氢颗粒污泥从蔗糖中产氢的影响

This work evaluated the effects of sodium ion concentration, ranging from 0 to 16000mg·L-1(Na ), on the conversion of sucrose to hydrogen by a high-activity anaerobic hydrogen-producing granular sludge. At the optimum sodium ion concentration [1000-2000mg·L-1(Na )] for hydrogen production at 37℃, the maximum sucrose degradation rate, the specific hydrogen production yield and the specific hydrogen production rate were 393.6-413.1mg·L-1·h-1, 28.04-28.97ml·g-1, 7.52-7.83ml·g-1·h-1, respectively. The specific production yields of propionate, butyrate and valerate decreased, with increasing sodium ion concentration, whereas the specific acetate production yield increased, meanwhile the specific production yields of ethanol and caproate were less than 55.3 and 12.6mg·g-1, respectively. The hybrid fermentation composition gradually developed from acetate, propionate and butyrate to acetate with the increase in sodium ion concentration.     

三段式谷氨酸棒杆菌发酵生产生物絮凝剂的过程及动力学模拟(英文)

Fermentation of bioflocculant with Corynebacterium glutamicum was studied by way of kinetic modeling. Lorentzian modified Logistic model, time-corrected Luedeking–Piret and Luedeking–Piret type models were pro-posed and applied to describe the cell growth, bioflocculant synthesis and consumption of substrates, with the correlation of initial biomass concentration and initial glucose concentration, respectively. The results showed that these models could well characterize the batch culture process of C. glutamicum at various initial glucose con-centrations from 10.0 to 17.5 g·L?1. The initial biomass concentration could shorten the lag time of cel growth, while the maximum biomass concentration was achieved only at the optimal initial glucose concentration of 16.22 g·L?1. A novel three-stage fed-batch strategy for bioflocculant production was developed based on the model prediction, in which the lag phase, quick biomass growth and bioflocculant production stages were sequentially proceeded with the adjustment of glucose concentration and dissolved oxygen. Biomass of 2.23 g·L?1 was obtained and bioflocculant concentration was enhanced to 176.32 mg·L?1, 18.62% and 403.63%higher than those in the batch process, respectively, indicating an efficient fed-batch culture strategy for bioflocculant production.     

丙酸盐影响利迪链霉菌AS 4.2501生产利迪链菌素及其碳流分布

To achieve higher antibiotic streptolydigin productivity through metabolic regulation, propionate was fed during the fermentation of Streptomyces lydicus AS 4.2501. The effects of propionate feeding on streptolydigin production and intracellular fluxes were investigated. The highest streptolydigin production yield of 95.10mg·L^-1 was obtained when 2mmol·L^-1 of sodium propionate was added at 60h of cultivation into shake-flask culture. This yield is 23.06% higher when compared to that of a batch culture without propionate supplementation. It was also found that when propionate was added, much more organic acids were excreted. Metabolic flux analysis was performed and it demonstrated that the carbon fluxes of the pentose phosphate pathway and the anaplerotic reaction were significantly increased after propionate feeding. The carbon flux from pyruvate to acetyl-CoA was determined to be 24.7, which was 12.27% higher than that in the batch culture. This study indicated that the glucose-6-phosphate and pyruvate nodes were potential bottlenecks for increasing streptolydigin productivity. Potential targets and strategies that could be manipulated through genetic and process engineering to increase the production of streptolydigin were also suggested.     

重组大肠杆菌流加发酵中短暂溶氧休克促进5-氨基乙酰丙酸生成(英文)

5-Aminolevulinic acid （ALA） is a common precursor for tetrapyrrole compounds in all kinds of organ isms and has wide applications in agriculture and medicines. In this study, a new strategy, i.e. short-term dissolved oxygen （DO） shock during aerobic fermentation, was introduced to produce 5-aminolevulinic acid with a recombi-nant E. coli. Effects of duration time of DO shock operation on plasmid concentration, intracellular ALA synthase （ALAS） activity and ALA production were investigated in Erlenmeyer shake flasks. The results indicated that both ALAS activity and ALA yield were enhanced in an anaerobic operation of 45 rain in the early exponential phase during fermentation, while they decreased when the anaerobic operation time was further increased to 60 rain. The DO shock protocol was confirmed with the fed-batch fermentation in a 15 L fermenter and the ALA production achieved 9.4 g.L-1 （72 mmol.L-1）, which is the highest yield in the fermentation broth reported up to now.     

Anaerobic hydrogen production of molasses from mixed microbial communities immobilized by activated granular carbon

Molasses wastewater was evaluated as substrate for biohydrogen production by anaerobic fermentation in a novel continuous mixed attached growth reactor ( CMAGR ) with aeration pretreated sludge attached onto granular activated carbon under continuous flow condition.It was indicated that the CMAGR system was operated at the conditions of influent COD of 2000～6000mg / L , hydraulic retention time ( HRT ) of 6hand temperature of 35 ℃ , when the pH value and oxidation-reduction potential ( ORP ) ranged from 4.16and-434 mV respectively , stable ethanol-type fermentation was formed with the sum of ethanol and acetate concentration ratio of 89.3%to the total liquid products after 40days operation.The H 2 content in biogas and chemical oxygen demand ( COD ) removal were estimated to be 46.6% and 13% , respectively.It was also investigated that the effects of organic loading rates ( OLRs ) on CMAGR hydrogen production system.It was found that hydrogen production yield increased from 3.72 mmol / hL to 12.51 mmol / hL as OLRs increased from 8 kg / m 3 d to 32 kg / m 3 d.The maximum hydrogen production rate of 12.51mmol / hL at a OLR of 32kg / m 3 d and the maximum hydrogen yield by substrate consumed was 130.57 mmol / mol happened at OLR of 16 kg / m 3 d.Greater pHs appeared to be favour to butyrate production and the maximum of 0.51mol / mol was obtained at pH of 4.14.However , ethanol / acetate ratio was greater than 1.1at pH fluctuated between 3.4 - 3.6and 4.1 - 4.4which indicated that these pHs were favour to ethanol type fermentation.Therefore , the continuous mixed attached growth reactor ( CMAGR ) could be a promising attached growth system for biohydrogen fermentation.     

面包酵母No.6催化不对称合成(2S,5S)-2,5-已二醇反应特性

Baker’s yeast number 6 was selected by screening. It showed good catalytic activity and enantioselectivity for asymmetric reduction of 2,5-hexanedione to produce (2S,5S)-2,5-hexanediol. Gas chromatography-mass spectrometry (GC-MS) revealed that the intermediate was (S)-5-hydroxyhexane-2-one. Reduction of 2,5-hexanedione proceeded in a two-step reaction. The hydroxyketone was initially formed, and this intermediate was further re-duced to the diol. Factors influencing the product yield and the enantiomeric excess of the reduction of 2,5-hexandione catalyzed by baker’s yeast number 6 were investigated. Higher concentration (≤100 mmol&#8226;L－1) of 2,5-hexandione did not influence 5-hydroxyhexane-2-one production, but 2,5-hexanediol production was inhibited by excess accumulation (＞30 mmol&#8226;L－1) of intermediate. The optimal conditions were glucose as the co-substrate at an initial glucose concentration of 20 g&#8226;L－1, 34&#61616;C, pH 7.0 and cell concentration 60 g&#8226;L－1 (cell dry mass). Under the optimal condition and an initial substrate concentration of 30 mmol&#8226;L－1, the yield of 2,5-hexandiol was 78.7% and the enantiomeric excess of (2S,5S)-2,5-hexandiol was 94.4% for 24-h reduction.     

两步酶法合成阿糖鸟苷

The chemical synthesis of Guanine arabinoside （ara-G） is extremely complex, time-consuming, and seriously polluted. A two-step enzymatic synthesis process was developed to acquire ara-G easily. 2,6-Diaminopurine arabinoside （ara-DA） was first synthesized with purine nucleoside phosphorylase and pyrimidine nucleoside phosphorylase produced by Enterobacter aerogenes DGW-07. The conversion yield of ara-DA could reach above 90% when the reaction liquid contained 30 mmol·L^-1 uracil arabinoside as arabinose donor, 10 mmol·L^- 1 2,6-diaminopurine as arabinose acceptor in pH 7.0 20 mmol·L^-1 phosphate buffer, and reacted at 60℃ for 48h. Then, ara-DA was effectively transformed into ara-G with adenylate deaminase produced by Aspergillus oryzae DAW-01. The total process had no complex separation and purification.     

重组大肠杆菌分批-补料高密度发酵生产类人胶原蛋白的动力学

The kinetics of batch and fed-batch cultures of recombinant Escherichia coli producing human-like collagen was investigated. In the batch culture, a kinetic model of a simple growth-association system was concluded without consideration of cell endogeneous metabolism. The cell lag time, the maximum specific growth rate and Yx/s were determined as 1.75h, 0.65h^-1 and 0.51g·g^-1, respectively. In the fed-batch culture, different specific growth rates were set at （0.15, 0.2, 0.25h^-1） by the method of pseudo-exponential feeding, and the expressions for the specific rate of substrate consumption, the growth kinetics and the product formation kinetics of each phase were obtained. The result shows that the concentrations of cell and product can reach 77.5g·L^-1 and 10.2g·L^-1 respectively. The modal predictions are in good agreement with the experimental data.     

Pt recovery using Cyphos IL-101 immobilized in biopolymer capsules

Cyphos^® IL-101, a tetraalkylphosphonium chloride salt (ionic liquid, IL) has been immobilized in capsules prepared by ionotropic gelation in calcium chloride solutions. The IL content was varied in the resin between 0.29 and 1.28 mmol IL g⁻¹. These resins have been tested for Pt recovery from HCl solutions. The equilibrium was very slightly affected by the concentration of HCl and chloride ions. The sorption isotherms were modeled using the Langmuir equation: the maximum sorption capacity was influenced by the drying of the resin but remained close to 177 mg Pt g⁻¹ for wet resin (i.e. 0.9 mmol Pt g⁻¹, dry weight basis, or 0.7 mol Pt mol⁻¹ Cyphos) and 142 mg Pt g⁻¹ for dry resin (i.e. 0.73 mmol Pt g⁻¹, or 0.57 mol Pt mol⁻¹ Cyphos). The presence of nitrates, nickel or copper ions (added under the form of chloride salts) did not significantly decrease sorption capacity even at concentrations as high as 5 g L⁻¹. Conversely, zinc at the concentration of 5 g L⁻¹, significantly decreased Pt sorption, probably due to the competition effect of chloro-anionic Zn species. This is another evidence of the ion exchange mechanism involved in the binding of hexachloroplatinate species. The kinetics are weakly affected by the agitation speed (in the range 150–350 rpm) indicating that the resistance to film diffusion is not the limiting step. The kinetics are affected by the IL content, metal concentration and more specifically the drying of the resin: intraparticle diffusion sounds to be the controlling kinetic step: the intraparticle diffusion coefficient varied between 2 × 10⁻¹² and 18 × 10⁻¹¹ m² min⁻¹, depending on experimental conditions. Platinum can be desorbed from loaded resin using either nitric acid (5 M) or thiourea (0.1 M in 0.1 M HCl acid solution). The resin was efficiently used for three sorption/desorption cycles using nitric acid: a decrease in sorption capacity and desorption efficiency was observed beginning with the third cycle, probably due to a progressive degradation of the resin.     

Selective precipitation of cadmium from nickel cadmium sulphate solutions using sodium decanoate

Each year in France, about 1300 tons of spent Ni–Cd batteries are collected. Their treatment consists of crushing, physical separation, acidic leaching of electrode materials, separation and recovery of metals leached with different processes. In this work, the selective precipitation of cadmium in synthetic Ni–Cd mixtures was investigated using sodium decanoate as precipitant. A factorial design of experiments was used to improve the separation using variables such as cadmium concentration, pH, molar ratio between decanoate and cadmium, time of addition of precipitant and rest time of the solution. In the best conditions defined by a 2⁵⁻² fractional factorial design, starting with concentrations of 0.1 mol L⁻¹ of nickel (5.9 g L⁻¹) and 0.05–0.15 mol L⁻¹ of cadmium (5.6–16.8 g L⁻¹) it is possible to recover in only one stage about 99% of cadmium without precipitating more than 5% of nickel. The results of the separation are on one side a pure solution of nickel at 2.39 × 10⁻² mol L⁻¹ (1.4 g L⁻¹) containing 3.44 × 10⁻⁴ mol L⁻¹ of cadmium (about 40 mg L⁻¹) and on the other side a precipitate of cadmium decanoate with a cadmium content equal to 23.6 wt% and nickel content lower than 0.8 wt%. These results demonstrated the viability of this separation.     

Antioxidant and Antimicrobial Activities of Echinacea (Echinacea purpurea L.) Extracts Obtained by Classical and Ultrasound Extraction

Antioxidant and antimicrobial activities of Echinacea purpurea L. (Asteraceae) extracts obtained by classical and ultrasound solvent extraction were compared. The dry aerial part of plant was extracted by 70% ethanol at a solid-to-liquid ratio of 1:10 (m/v) and 25°C. The extract obtained by classical solvent extraction contained 29% larger amounts of phenolic compounds and 20% higher content of flavonoids. 2,2-diphenyl-1-picril hydrazyl radical (DPPH) scavenging reached 93.6% and the values of EC50 were (34.16±0.65) μg·ml⁻¹ and (65.48±1.12) μg·ml⁻¹ for the extracts obtained by the classical and ultrasound extractions, respectively. The extracts, independent of the extraction technique applied, showed a considerable growth inhibition on Candida albicans and Saccharomyces cerevisiae, while no growth inhibition zones were observed for Aspergillus niger. The diameters of inhibition zone observed for all the microorganisms were larger for extracts obtained by classical extraction than those by ultrasound extraction.     

Valorisation of hardwood hemicelluloses in the kraft pulping process by using an integrated biorefinery concept

A primary hydrolysis treatment (auto or acid-catalysed) of Eucalyptus globulus wood was performed before the cooking stage to extract part of the hemicelluloses that otherwise would be dissolved in the kraft liquor and burned. As xylose was the main monosaccharide Pichia stipitis was selected to produce bioethanol. Two methods were tested, with different alkalis, to reduce hydrolysates toxicity and adjust pH. A two-step method using Ca(OH)₂ leads to better fermentation results. Acid hydrolysates promoted higher ethanol concentrations (12 g L⁻¹) with high productivity and yield values (0.22 g_eth L⁻¹h⁻¹ and 0.48 g_eth/g_{xyl eqs}), whilst auto-hydrolysates, even after a secondary hydrolysis, gave low ethanol concentrations (2–4 g L⁻¹). The impacts on kraft cooking and pulp quality were also studied in order to fully understand the feasibility of this biorefinery concept (combining ethanol production and hardwood pulping). A decrease of the overall pulp yield (10% for auto- and 15% for acid-hydrolysis) was observed. However, a decrease on bleaching requirements (up to 15%) and on brightness reversion was registered. Moreover, auto-hydrolysis improves pulp viscosity, whilst acid-hydrolysis is more efficient in metals leaching. Overall, from the pulp production point of view, auto-hydrolysis conditions are more attractive than the acid-catalysed ones.     

一种新型反应器——生物气射流循环厌氧流化床的启动与运行特性

A novel anaerobic reactor, jet biogas inter-loop anaerobic fluidized bed （JBILAFB）, was designed and constructed. The start-up and performance of the reactor was investigated in the Process. of .artificial glucose wastewater treatment. With the wastewater recycle ratio of 2.5 ： 1, the recycled wastewater with biogas could mix sludge and wastewater in the JBILAFB reactor completely. The start-up of the JBILAFB reactor could be completed in less than 70 d through maintenance of hydraulic retention time （HR~I＂） and stepwise increase of feed total organic carbon （TOC） concentration. After the start-up, with the volumetric TOC loadings of 14.3 kg·m ^-3·d^-1, the TOC removal ratio, the effluent pH, and the volatile fatty acids （VFA）/alkalinity of the JBILAFB reactor were more than 80%, close to 7.0 and less than 0.4, respectively. Moreover, CH4 was produced at more than 70% of the theoretical value, The reactor exhibited high stability under the condition of high volumetric TOC loading. Sludge granules in the JBILAFB reactor were developed during the start-up and their sizes were enlarged with the stepwise increase of volumetric TOC loadings from 0.8 kg.m^-3.d ^-1 to 14.3 kg.m^-3.d^-1. Granules, an offwhite color and a similar spherical shape, were mainly comprised of global-like bacteria. These had good methanogenic activity and settleability, which were formed probably through adhesion of the bacteria. Some inorganic metal compounds such as Fe, Ca, Mg, Al, etc. were advantageous to the formation of the granules.     

Electrochemical diamond sensors for TNT detection in water

An electrochemically stabilized boron doped diamond electrode prepared by chemical vapour deposition (CVD) is used for electrochemical TNT sensing in aqueous solutions. Square wave voltammograms (SWVs) exhibit three highly resolved peaks at −0.47, −0.62 and −0.76 V vs. Ag–AgCl reference electrode, respectively. The current vs. TNT concentration plot shows a linear relationship with a same slope for the two first TNT peaks at μg L⁻¹ and mg L⁻¹ concentration ranges. Detection and quantification limits of 10 and 25 μg L⁻¹, respectively, were obtained without any preconcentration step. Relative standard deviation (RSD) of less than 1% measured over 10 runs has been found for the −0.47 V peak current showing the very high stability of the electrode without any significant fouling effect. An interference study with nitro aromatic compounds of the same family (nitro toluene and dinitrotoluene) has shown that the −0.47 V reduction peak enables TNT discrimination. Measurement of TNT in a natural medium (sea water without any purification step except filtering) has been also investigated.